Oral preparation useful in measuring capacity to metabolize pyridine

ABSTRACT

An object of the present invention is to provide an oral preparation that can be used to diagnose the existence or degree of pyridine metabolic capacity disorder, pyrimidine metabolic rate, etc., with high accuracy and with little variation due to individual differences. The oral preparation is prepared using a powder material obtained by mixing and pulverizing (a) an isotope-labeled compound and/or a pyrimidine metabolite compound and (b) a sugar and/or a sugar alcohol.

TECHNICAL FIELD

The present invention relates to an oral preparation that can beeffectively used to assess, with high accuracy, the existence or degreeof a pyridine metabolic capacity disorder; pyrimidine-metabolic rate;etc. The present invention also relates to a process for producing theoral preparation.

BACKGROUND ART

5-Fluorouracil (hereinafter sometimes referred to as “5-FU”), variousderivatives thereof (such as tegafur, carmofur, doxifluridine, etc.),and like fluorouracil drugs are widely used as anticancer drugs atpresent. It is known that 5-FU administered to the body is firstdegraded by the action of dihydropyrimidine dehydrogenase (hereinaftersometimes referred to as “DPD”), which is the first enzyme in thepyrimidine metabolic pathway. It is therefore believed that theconcomitant use of a drug that inhibits DPD enzymatic activity iseffective in sustaining the effects of fluorouracil drugs such as 5-FUand the like. On the other hand, it is known that when a fluorouracildrug such as 5-FU is administered to a subject with DPD deficiency orreduced DPD activity, the drug is not metabolized in a normal manner andresults in an abnormally high fluorouracil drug concentration in theblood, thereby causing severe side effects (e.g., myelosuppression,digestive symptoms, etc).

Thus, in order to effectively exhibit the action of fluorouracil drugsor prevent the side effects of fluorouracil drugs, diagnosis ofpyrimidine-metabolic capacity, i.e., the existence, degree, etc., of apyrimidine metabolic disorder in the subject, before administration of afluorouracil drug is believed to be important.

A method for diagnosing pyrimidine metabolic activity in a subject hasbeen reported in which an isotope-labeled pyrimidine compound isadministered to the subject, and the excretion behavior of theisotope-labeled metabolic product discharged from the body is measuredso as to determine the pyrimidine metabolic capacity, i.e., theexistence, degree, etc., of a pyrimidine metabolic disorder in thesubject (e.g., Patent Document 1). Granules and subtle granulescontaining isotope-labeled pyrimidine compounds and carriers are alreadyknown as pyrimidine metabolic capacity diagnosis preparations for use inthe above method.

However, isotope-labeled pyrimidine compounds, such as ¹³C-uracil, have,as well as low solubility, characteristically high cohesiveness,although bulk powders of such compounds themselves are fine particles ofseveral microns. Therefore, granules and subtle granules prepared fromisotope-labeled pyrimidine compounds as such by standard methods do notrapidly dissolve, and partly because of this, the compounds havedisadvantages such as a slow and non-uniform absorption rate in theliving body and variation in the absorption rate due to individualdifferences. Therefore, in order to realize pyrimidine metaboliccapacity diagnosis with higher accuracy, it is desired to overcome theabove defects so that variation in the excretion time and amount of theisotope-labeled metabolic products can be reduced and the non-uniformityof diagnosis accuracy due to individual differences can be decreased.

Patent Document 1: International Publication No. WO 02/072153, pamphlet

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention

An object of the present invention is to provide an oral preparationthat can be used to diagnose the existence, degree, etc., of a pyridinemetabolic capacity disorder, with high accuracy and with littlevariation due to individual differences.

Means for Solving the Problems

The present inventors conducted extensive research to solve the aboveproblems, and found that an oral preparation prepared using a powdermaterial obtained by mixing and pulverizing (a) an isotope-labeledcompound and/or a metabolite thereof, and (b) a sugar and/or a sugaralcohol, enables pyrimidine metabolic capacity diagnosis with highaccuracy and with little variation due to individual differences. Thepresent invention has been achieved by further improvements based onthis finding.

The present invention provides the following oral preparations,production processes for the same, etc.

Item 1. An oral preparation produced from a powder material obtained bymixing and pulverizing (a) a pyrimidine compound and/or a metabolitethereof, in which at least one of carbon atoms, oxygen atoms, andnitrogen atoms is labeled with an isotope, and (b) a sugar and/or asugar alcohol.

Item 2. An oral preparation according to item 1, wherein a particlediameter at 50% of the powder material is 40 μm or less.

Item 3. An oral preparation according to item 1, which contains thecomponent (a) in a proportion of 5 to 20 wt. %.

Item 4. An oral preparation according to item 1, wherein the component(a) is isotope-labeled uracil.

Item 5. An oral preparation according to item 1, wherein the component(b) is mannitol.

Item 6. An oral preparation according to item 1, wherein the component(a) is isotope-labeled uracil and the component (b) is mannitol.

Item 7. An oral preparation according to item 1, which is a granularpreparation.

Item 8. An oral preparation according to item 7, which is produced byextrusion granulation of the powder material.

Item 9. An oral preparation according to item 7, wherein the granularpreparation has a mean particle diameter of 1400 μm or less.

Item 10. An oral preparation according to item 1, which is a preparationfor diagnosing pyrimidine-metabolic capacity.

Item 11. An oral preparation according to item 1, which is a preparationfor determining gastric emptying capacity.

Item 12. An oral preparation according to item 1, which is a preparationfor diagnosing dyspepsia.

Item 13. A process for producing an oral preparation, the processcomprising the steps of:

(1) producing a powder material by mixing and pulverizing (a) apyrimidine compound and/or a metabolite thereof, in which at least oneof carbon atoms, oxygen atoms, and nitrogen atoms is labeled with anisotope, and (b) a sugar and/or a sugar alcohol; and

(2) formulating the powder material obtained in the above step (1) intoa preparation.

Item 14. A process according to item 13, wherein a particle diameter ofthe powder material produced in the step (1) is of 40 μm or less.

Item 15. A process according to item 13, wherein the oral preparationcontains the component (a) in a proportion of 5 to 20 wt. %.

Item 16. A process according to item 13, wherein the component (a) isisotope-labeled uracil.

Item 17. A process according to item 13, wherein the component (b) ismannitol.

Item 18. A process according to item 13, wherein the component (a) isisotope-labeled uracil and the component (b) is mannitol.

Item 19. A process according to item 13, wherein the oral preparationhas a granular form.

Item 20. A process according to item 19, wherein the step (2) is a stepof formulating the powder material obtained in the step (1) into apreparation by extrusion granulation.

Item 21. A process according to item 19, wherein the oral preparation isa granular preparation having a mean particle diameter of 1400 μm orless.

Item 22. A process according to item 13, wherein the oral preparation isa preparation for diagnosing pyrimidine metabolic capacity.

Item 23. A process according to item 13, wherein the oral preparation isa preparation for determining gastric emptying capacity.

Item 24. A process according to item 13, wherein the oral preparation isa preparation for diagnosing dyspepsia.

Item 25. Use of a powder material obtained by mixing and pulverizing (a)a pyrimidine compound and/or a metabolite thereof, in which at least oneof carbon atoms, oxygen atoms, and nitrogen atoms is labeled with anisotope, and (b) a sugar and/or a sugar alcohol, for producing apreparation for diagnosing pyrimidine metabolic capacity.

Item 26. Use of a powder material obtained by mixing and pulverizing (a)a pyrimidine compound and/or a metabolite thereof, in which at least oneof carbon atoms, oxygen atoms, and nitrogen atoms is labeled with anisotope, and (b) a sugar and/or a sugar alcohol, for producing apreparation for determining gastric emptying capacity.

Item 27. Use of a powder material obtained by mixing and pulverizing (a)a pyrimidine compound and/or a metabolite thereof, in which at least oneof carbon atoms, oxygen atoms, and nitrogen atoms is labeled with anisotope, and (b) a sugar and/or a sugar alcohol, for producing apreparation for diagnosing dyspepsia.

Effects of the Invention

The oral preparation of the present invention is produced by formulatinga powder material obtained by mixing and pulverizing (a) anisotope-labeled compound and/or a metabolite thereof and (b) a sugarand/or a sugar alcohol, into a preparation. With such formulation, theoral preparation of the present invention makes it possible to diagnosepyrimidine metabolic capacity and gastric emptying capacity with highaccuracy and with little variation due to individual differences. As aresult, the behavior of an isotope-labeled metabolic product can becorrectly determined by one or a small number of measurements, 20 to 30minutes after administration of the preparation, so that the timerequired for the determination and the number of measurements can bereduced, thereby decreasing the burden on patients.

BRIEF DESCRIPTION OF THE DRAWINGS

[FIG. 1]A figure showing the behavior of degradation (metabolism) ofpyrimidine compounds (uracil, 5-fluorouracil (5-FU), and thymine) by aseries of pyrimidine metabolizing enzymes (dihydropyrimidinedehydrogenase (DPD), dihydropyrimidinase (DHPase), andβ-ureidopropionase (β-UPase)).

[FIG. 2]A figure comparing the results of observing, over time, thebehavior of ¹³CO₂ excreted in the expired air of three healthy subjects(Subjects A, B, and C) to whom the granular preparation of Example 2 hasbeen administered.

[FIG. 3]A figure comparing the results of observing, over time, thebehavior of ¹³CO₂ excreted in the expired air of three healthy subjects(Subjects A, B, and C) to whom the granular preparation of ComparativeExample 2 has been administered.

[FIG. 4]A figure-showing, over time, the behavior of ¹³CO₂ excreted inthe expired air of 20 patients suspected of having gastroparesis, towhom the preparation of Example 1 has been administered in Test Example5.

[FIG. 5]A figure showing the plasma 2-¹³C uracil concentration inpatients divided into three groups (normal gastric emptying capacity,reduced gastric emptying capacity, and insufficient gastric emptyingcapacity) based on the results shown in FIG. 4, 20 minutes afteradministration of the preparation of Example 1.

BEST MODE FOR CARRYING OUT THE INVENTION

The present invention is described below in detail.

The oral preparation of the present invention contains anisotope-labeled pyrimidine compound and/or a metabolite thereof(hereinafter these are sometimes referred to as “Component (a)”).

The pyrimidine compound for use in the present invention may be any of awide variety of compounds having a pyrimidine skeleton, and ispreferably a compound that serves as a substrate for a pyrimidinemetabolizing enzyme, and in particular dihydropyrimidine dehydrogenase(DPD), which is the first enzyme in the pyrimidine metabolic pathway inthe living body. Specific examples of such pyrimidine compounds includeuracil, thymine, and derivatives thereof. The derivatives of uracil andthymine are not limited as long as they serve as substrates for DPD, andas long as their final metabolic products formed via the pyrimidinemetabolic pathway are discharged in excrement such as expired air,urine, or sweat. Specific examples of such derivatives include halidesof uracil, such as 5-fluorouracil, 5-bromouracil, etc.; halides ofthymine, such as 5-fluorothymine, 5-bromothymine, etc.; and the like.Preferable examples of pyrimidine compounds include uracil, thymine, and5-fluorouracil.

Usable pyrimidine compounds include, in addition to the above compounds,which serve as direct substrates for DPD, compounds that serve asindirect substrates for the enzyme, i.e., precursors (includingprodrugs), which are metabolized or degraded in vivo into substrates forDPD (such as uracil, thymine, 5-fluorouracil, etc.). Examples of suchprecursors include precursors of uracil, such as cytosine, uridine, andphosphates thereof (e.g., uridylic acid); precursors of thymine, such as5-methylcytosine, thymidine, and phosphates thereof (e.g., thymidylicacid); and precursors (prodrugs) of 5-fluorouracil, such as tegafur,carmofur, doxifluridine, etc.

The metabolite of a pyrimidine compound is a compound that correspondsto a metabolic intermediate of the pyrimidine compound and that servesas a substrate for a pyrimidine metabolizing enzyme, and in particulardihydropyrimidinase (hereinafter sometimes referred to as “DHPase”),which is the second enzyme in the pyrimidine metabolic pathway in theliving body, or β-ureidopropionase (hereinafter sometimes' referred toas “β-UPase”), which is the third enzyme. Specific examples ofmetabolites of pyrimidine compounds include dihydrouracil,dihydrothymine, and derivatives thereof (e.g., halides of dihydrouracil,such as 5-fluorodihydrouracil and the like), which serve as substratesfor DHPase; and β-ureidopropionic acid, β-ureidoisobutyric acid, andderivatives thereof (e.g., halides of β-ureidopropionic acid, such asfluoro-β-ureidopropionic acid, and halides of β-ureidoisobutyric acid),which serve as substrates for β-UPase.

In the present invention, Component (a) is preferably a pyrimidinecompound, more preferably uracil, thymine, or 5-fluorouracil, and stillmore preferably 5-fluorouracil.

In the pyrimidine compound and/or metabolite thereof for use in thepresent invention, at least one of the carbon atoms, oxygen atoms, andnitrogen atoms in the molecule is labeled with an isotope. The isotopeis not limited, and specific examples include ¹³C, ¹⁴C, ¹⁸O, and ¹⁵N.The isotope may be radioactive or non-radioactive, but ¹³C, ¹⁸O, or ¹⁵N,which are non-radioactive, are preferable from the viewpoint of safety.

The pyrimidine compound and/or metabolite thereof for use in the presentinvention may have one isotope in the molecule or may have two or moreisotopes of the same or different elements. Although not limitative, itis preferable that a carbon atom or oxygen atom in the pyrimidinecompound or metabolite thereof be labeled so that at least part (C or O)of CO₂ produced via the pyrimidine metabolic pathway is labeled with anisotope. Examples of such pyrimidine compounds include those having anisotope-labeled carbon atom at the 2-position of the pyrimidineskeleton. Specific examples include 2-¹³C-labeled uracil and2-¹³C-labeled fluorouracil.

The method for labeling a pyrimidine compound and/or a metabolitethereof with an isotope as mentioned above is not limited, and a widevariety of conventional methods can be employed (Sasaki, “5.1Application of Stable Isotopes in Clinical Diagnosis”; Kagaku no Ryoiki(Journal of Japanese Chemistry) 107, “Application of Stable Isotopes inMedicine, Pharmacy, and Biology”, Nankodo, pp. 149-163 (1975); Kajiwara,“RADIOISOTOPES”, 41, 45-48 (1992); etc.). Some of such isotope-labeledpyrimidine compounds and metabolites thereof are commercially available,and these commercial products are conveniently usable.

The proportion of Component (a) in the oral preparation of the presentinvention is, for example, usually 5 to 20 wt. %, preferably 6 to 18 wt.%, and more preferably 8 to 15 wt. %.

The oral preparation of the present invention contains, in addition toComponent (a), a sugar and/or a sugar alcohol (hereinafter these aresometimes referred to as “Component (b)”).

The sugar for use in the present invention is not limited as long as itis pharmaceutically acceptable. Examples of such sugars include glucose,galactose, fructose, xylose, arabinose, mannose, and likemonosaccharides; maltose, isomaltose, cellobiose, lactose, sucrose,trehalose, and like disaccharides; etc. Among these, glucose and sucroseare preferable.

The sugar alcohol for use in the present invention is not limited aslong as it is pharmaceutically acceptable. Specific examples of sugaralcohols include erythritol, mannitol, xylitol, sorbitol, maltitol,reducing paratinose, lactitol, etc. Among these, mannitol, xylitol, anderythritol are preferable, and mannitol is more preferable.

In the present invention, Component (b) is preferably a sugar alcohol,more preferably mannitol, xylitol, or erythritol, and still morepreferably mannitol.

The proportion of Component (b) in the oral preparation of the presentinvention is, for example, usually 80 to 95 wt. %, preferably 82 to 94wt. %, and more preferably 85 to 92 wt. %, based on the total weight ofthe preparation.

The ratio of Component (b) to Component (a) in the oral preparation ofthe present invention is, for example, 400 to 1900 parts by weight,preferably 450 to 1550 parts by weight, and more preferably 550 to 1150parts by weight, of Component (b), per 100 parts by weight of Component(a). The combined use of Components (a) and (b) in such a ratio furtherimproves the accuracy of pyrimidine metabolic disorder diagnosis.

The oral preparation of the present invention is produced by formulatinga powder material containing Components (a) and (b) into a preparation.The powder material used for preparing the oral preparation of thepresent invention is obtained by mixing Components (a) and (b) in theabove ratio and pulverizing the resulting mixture.

The oral preparation of the present invention may have the samecomposition as the powder material after pulverization, or may containother components in addition to the powder material. Therefore, theproportions of Components (a) and (b) in the powder material aresuitably selected according to the proportions of Components (a) and (b)in the final form of the oral preparation, the preparation steps for theoral preparation, etc.

The powder material may be obtained by mixing and pulverizingpharmaceutically acceptable additives together with Components (a) and(b), as long as the effects of the present invention are not impaired.Such additives are the same as those that can be added when formulatingthe powder material into a preparation. Specific examples of suchadditives are given hereinafter.

The particle diameter of the powder material is not limited as long asthe particle diameter is a result from mixing and pulverizing Components(a) and (b), but in order to increase the accuracy of pyrimidinemetabolic capacity diagnosis, it is desirable that the particle diameterat 50% be 40 μm or less, preferably 30 μm or less, and more preferably 5to 20 μm.

Preferable examples of the powder material are powder materials having aparticle size distribution such that the particle diameter at 50% is 40μm or less and the particle diameter at 90% is 200 Cm or less; morepreferable examples are those having a particle size distribution suchthat the particle diameter at 50% is 30 μm or less, and the particlediameter at 90% is 100 μm or less; and still more preferable examplesare those having a particle size distribution such that the particlediameter at 50% is 5 to 20 μm, and the particle diameter at 90% is 10 to70 μm. Use of a powder material having such particle size distributionto prepare the oral preparation enables Component (a) to be absorbed inthe living body at a rapid and uniform rate, thereby making it possibleto diagnose pyrimidine metabolic capacity with higher accuracy.

As used herein, the meanings of the particle diameter at 50% andparticle diameter at 90% of the powder material are as follows: thevolume of the particles of the powder material is integrated in orderfrom the particle with the smallest particle diameter, until theintegrated volume accounts for 50% or 90% of the total volume of theparticles of the powder material, and the particle diameter of the lastparticle integrated is the particle diameter at 50% or 90%. The particlediameter at 50% and particle diameter at 90% can be measured using a drylaser method (measurement conditions: a focal distance of 100 mm, anumber of averaging processes of 10, an averaging interval of 5milliseconds, and an air pressure of 0.4 MPa).

The pulverizing treatment used for the preparation of the powdermaterial is not limited, but pulverizing treatment using a dry mill ispreferable. Specific examples of dry mills include hammer mills, pinmills, jet mills, etc.

The oral preparation of the present invention is produced by adding, asrequired, additives such as excipients, binders, pH adjusters,disintegrators, absorption enhancers, lubricants, colorants, corrigents,flavors, etc., to the powder material, and formulating the resultingmixture into a preparation via a treatment such as granulation oranother forming procedure, which is selected according to the form ofthe preparation. When the oral preparation of the present invention is apowder preparation, the powder material as such can be used as the oralpreparation in the final form.

Specific examples of additives that can be used for formulation includelactose, starch, refined white sugar, dextrin, mannitol, xylitol,sorbitol, erythritol, calcium dihydrogen phosphate, sodium chloride,glucose, calcium carbonate, kaolin, crystalline cellulose, silicate, andlike excipients; water, ethanol, simple syrup, glucose solutions, starchsolutions, gelatin solutions, carboxymethylcellulose,carboxymethylcellulose sodium, shellac, methylcellulose,hydroxypropylmethylcellulose, hydroxypropylcellulose,polyvinylpyrrolidone, polyvinyl alcohol, gelatin, dextrin, pullulan, andlike binders; citric acid, citric anhydride, sodium citrate, sodiumcitrate dihydrate, anhydrous sodium monohydrogen phosphate, anhydroussodium dihydrogen phosphate, sodium hydrogen phosphate, anhydrous sodiumdihydrogen phosphate, and like pH adjusters; carmellose calcium,low-substituted hydroxypropycellulose, carmellose, croscarmellosesodium, carboxymethyl starch sodium, crospovidone, and likedisintegrators; polysorbate 80, quaternary ammonium bases, sodium laurylsulfate, and like absorption enhancers; purified talc, stearate,polyethylene glycol, colloidal silicic acid, sucrose fatty acids,hydrogenated oils, and like lubricants; yellow iron oxide, yellow ironsesquioxide, iron sesquioxide, β-carotene, titanium oxide, food colors(e.g., Food Blue No. 1), copper chlorophyll, riboflavin, and likecolorants; ascorbic acid, aspartame, sweet hydrangea leaf, sodiumchloride, and like corrigents; and the like.

The form of the oral preparation of the present invention is not limitedas long as it is a solid preparation, and subtle granules, granules,powders, tablets (including naked tablets and coated tablets), capsules,pills, and other forms can be selected as desired. Among these, tofurther enhance the effects of the present invention, granularpreparations such as subtle granules and granules, and in particulargranular preparations produced by extrusion granulation, are preferred.

When the oral preparation of the present invention is a granularpreparation, the mean particle diameter of the preparation is, forexample, usually 1400 μm or less, preferably 50 to 1200 μm, and morepreferably 100 to 1000 μm. When the granular preparation has such aparticle diameter, the granular preparation enables pyrimidine metaboliccapacity diagnosis with higher accuracy. The particle diameter of thepreparation can be measured using a vibration sieve method(specifically, using a measurement apparatus Robot Shifter RPS-95(Seishin Enterprise Co., Ltd.) at a vibration level of 5, a shift timeof 5 minutes, and a pulse interval of 1 second).

After administering the oral preparation of the present invention, thepyrimidine metabolic capacity, i.e., the existence or degree of apyrimidine metabolic disorder, pyrimidine metabolic rate, etc., in asubject, can be assessed by measuring the excretion behavior of theisotope-labeled metabolic product excreted from the body. Therefore, theoral preparation of the present invention can be used as a preparationfor determining pyrimidine metabolic capacity. Further, as describedhereinafter, since gastric emptying capacity can also be assessed basedon the assessment results of pyrimidine metabolic capacity, andspecifically the results of measuring pyrimidine metabolic rate, theoral preparation of the present invention can also be used as apreparation for determining gastric emptying capacity. Embodiments ofthe preparation for determining pyrimidine metabolic capacity and thepreparation for determining gastric emptying capacity are specificallydescribed below.

Preparation for Determining Pyrimidine Metabolic Capacity

Since the oral preparation of the present invention can be used todetermine pyrimidine metabolic capacity with respect to the existence,degree, etc., of a pyrimidine metabolic disorder, the preparation isuseful for the detection, measurement, and diagnosis of a pyrimidinemetabolic disorder. Specific conditions, method, etc., for using theoral preparation of the present invention as a preparation fordetermining pyrimidine metabolic capacity are as follows.

When the oral preparation of the present invention is administered to asubject with normal pyrimidine metabolic capacity, in whom or which theseries of pyrimidine metabolizing enzymes (DPD, DPHase, and β-UPase)function normally in the living body (hereinafter sometimes referred toas a “healthy subject”), the pyrimidine compound contained as Component(a) in the preparation is metabolically degraded into metabolic productssuch as β-alanine, F-β-alanine, β-aminoisobutyric acid, NH₃, CO₂, etc.,as shown in FIG. 1.

The final metabolic product CO₂ thus formed by metabolism is excreted inexpired air, and β-alanine, F-β-alanine, or β-aminoisobutyric acid isexcreted mainly in urine. Of the final metabolic products thus excreted,at least one of CO₂ and a final metabolic product selected fromβ-alanine, F-β-alanine, and β-aminoisobutyric acid is labeled with anisotope, depending on the isotope-labeled site of the pyrimidinecompound and/or metabolite thereof used as Component (a). Such anisotope label is used as an index to measure the excretion behavior (thebehavior of excretion amount or excretion rate over time) of these finalmetabolic products using, as a test sample, expired air when CO₂ islabeled, or urine when β-alanine, F-β-alanine, β-aminoisobutyric acid,or ammonia is labeled.

The pyrimidine metabolic capacity of the subject can be determined fromthe thus measured excretion behavior (the behavior of excretion amountor excretion rate over time) of the isotope-labeled metabolic product.

When the oral preparation of the present invention is used fordetermining pyrimidine metabolic capacity, the dose of the oralpreparation of the present invention is not limited, but is preferablyan amount corresponding to 1 to 2000 mg, and preferably 10 to 300 mg, ofComponent (a).

When using the oral preparation of the present invention for determiningpyrimidine metabolic capacity, it is preferable to use as Component (a)a pyrimidine compound and/or a metabolite thereof that causesisotope-labeled CO₂ to be excreted in expired air as a result ofmetabolism. Using such a preparation, the pyrimidine metabolic capacityof a subject can be determined from the excretion behavior (the behaviorof excretion amount and excretion rate over time) of isotope-labeledCO₂, which can be found by administering the preparation to the subjectand measuring isotope-labeled CO₂ excreted in the expired air of thesubject.

When the preparation contains, as an active ingredient, a pyrimidinecompound that forms an isotope-labeled compound other thanisotope-labeled CO₂, such as β-alanine, fluoro-β-alanine,β-aminoisobutyric acid, or the like, excrement such as urine, sweat orthe like is used in place of expired air as a test sample.

When expired air is used as a test sample, the method for measuringisotope-labeled CO₂ contained in expired air varies depending on whetherthe isotope used is radioactive or non-radioactive. Conventionalanalytic methods are usable, including a liquid scintillation-countermethod, mass spectrometry, infrared spectrometry, emission spectrometry,magnetic resonance spectrometry, etc. From the viewpoint of measurementaccuracy, infrared spectrometry and mass spectrometry are preferable.When excrement such as urine, sweat, or the like is used as the testsample, the isotope-labeled pyrimidine compound (or an isotope-labeledpyrimidine metabolite), isotope-labeled metabolic intermediates, andisotope-labeled metabolic products contained in the test sample can beseparated simultaneously and analyzed at the same time by the combineduse of separation techniques, such as liquid chromatography, gaschromatography, etc. Thus, the excretion behavior of the isotope-labeledmetabolites can be selectively measured.

The pyrimidine metabolic capacity in a subject can be assessed by, forexample, comparing the excretion behavior (the behavior of the excretionamount or excretion rate over time) of an isotope-labeled metabolicproduct in the subject, which is measured as described above, with theexcretion behavior of the isotope-labeled metabolic product in a healthysubject having a normal pyrimidine metabolic capacity, which is measuredin the same manner. Specifically, when isotope-labeled CO₂ excreted inexpired air is measured as an isotope-labeled metabolic product, theamount of isotope-labeled CO₂ gas at a predetermined time afteradministration of the oral preparation, carbon dioxide gas Δ (%) value(difference in the isotope-labeled ¹³CO₂/¹²CO₂ concentration ratiobetween the expired air samples collected before and afteradministration of the oral preparation), or the initial rate ofisotope-labeled CO₂ gas excreted rate in expired air, can be used as anindex of the excretion behavior of the isotope-labeled metabolicproduct. For example, using the carbon dioxide gas Δ (%) value orinitial rate in a healthy subject as a standard, a subject having alower carbon dioxide gas Δ (%) value or lower initial rate is diagnosedas having reduced pyrimidine metabolic capacity.

Further, in place of or in addition to the excretion behavior of anisotope-labeled metabolic product, the area under the curve (AUC),excretion rate (especially the initial excretion rate), maximumexcretion concentration (Cmax), or like parameter, preferably apharmacokinetic parameter, in a test subject, can be compared with thecorresponding parameter in a healthy subject.

The deficiency or existence of a pyrimidine metabolizing enzyme (atleast one of DPD, DHPase, and β-UPase) can be determined based on theexistence or non-existence of the excretion of the isotope-labeledmetabolic product, without comparison with the excretion behavior of ahealthy subject. The existence of a decrease or increase in pyrimidinemetabolic capacity (pyrimidine metabolic disorder), and the degreethereof (degree of the disorder) can be determined by comparing theexcretion behavior in the subject or a parameter obtained therefrom,with the corresponding excretion behavior or parameter in a healthysubject.

Preparation for Determining Gastric Emptying Capacity

When using the oral preparation of the present invention for determininggastric emptying capacity, it is preferable to use as Component (a) apyrimidine compound and/or a metabolite thereof that causesisotope-labeled CO₂ to be excreted in expired air as a result ofmetabolism.

After being orally ingested by a subject, the oral preparation of thepresent invention enters the stomach, and is finally discharged throughthe pylorus by the contraction-relaxation and peristalsis of thestomach. After being discharged from the pylorus, Component (a) israpidly absorbed in the duodenum and lower parts of the gastrointestinaltract (the duodenum, jejunum, ileum, etc.), metabolized, and excreted inexpired air as isotope-labeled CO₂ gas. Component (a) used in the oralpreparation of the present invention is not at all or hardly absorbed inthe stomach, but after being discharged from the stomach, the componentis rapidly absorbed, metabolized, and excreted in expired air asisotope-labeled CO₂ gas. Therefore, the excretion behavior ofisotope-labeled CO₂ gas in expired air (expressed as, for example, aratio of isotope-labeled CO₂ gas relative to ¹²CO₂ excreted in theexpired air (isotope-labeled CO₂/¹²CO₂)) depends on the gastric emptyingrate (gastric emptying time) of Component (a) contained in the oralpreparation of the present invention.

The dose of the oral preparation of the present invention may be thesame as in the case where the oral preparation of the present inventionis used for determining pyrimidine metabolic capacity.

Isotope-labeled CO₂ contained in expired air can be measured using thesame method as in the case where the oral preparation of the presentinvention is used for determining pyrimidine metabolic capacity.

The gastric emptying capacity in a subject can be assessed using, as agastric emptying capacity index, the amount of isotope-labeled CO₂ gasat a predetermined time after administration of the oral preparation,the carbon dioxide gas Δ (%) value (difference in the isotope-labeledCO₂/¹²CO₂ concentration ratio between expired air samples collectedbefore and after administration of the oral preparation), or initialrate of isotope-labeled CO₂ gas excreted rate. For example, using thecarbon dioxide gas Δ (%) value or initial rate in a healthy subject as astandard, a subject having a lower carbon dioxide gas Δ (%) value orinitial rate can be diagnosed as having reduced gastric emptyingcapacity.

The oral preparation of the present invention can be administeredsingly, or may be administered at the same time as or immediately beforeor after ingestion of a test meal. Preferably, the gastric emptyingcapacity-determining composition of the present invention isadministered immediately after ingestion of a test meal. The test mealis not limited as long as it does not impair the effects of the gastricemptying capacity determination using the preparation of the presentinvention, and may be a solid food, fluid food, or liquid food.

The main cause of dyspepsia (non-ulcer upper gastrointestinal tractsyndrome) is a gastrointestinal motility disorder, and in particularreduction of gastric emptying capacity. Therefore, the oral preparationof the present invention can be effectively used as a preparation for adiagnostic test for dyspepsia, and in particular dyspepsia caused mainlyby insufficient gastric emptying capacity (e.g., dysmotility-likedyspepsia).

Further, use of the oral preparation of the present invention fordetermining gastric emptying capacity makes it possible to determine theefficacy, or the therapeutic effects on individual subjects, ofgastrointestinal drugs, and in particular drugs associated withgastrointestinal motor functions. Specifically, the determination can beperformed by measuring the gastric emptying capacity using the oralpreparation of the present invention before and after administration ofa gastrointestinal drug, and in particular a drug associated withgastric mobility function, and comparing the two measurements. Thisassesses the efficacy of the drug itself. In addition, since therapeuticeffects of a drug on individual subjects can also be assessed, the oralpreparation can also be used for selecting drugs that are suitable forindividual subjects. Examples of drugs associated with gastrointestinalmotor functions include drugs that control the peristalsis of thestomach by enhancement or suppression, such as gastrointestinal motorfunction improving agents, gastrointestinal motor function enhancers,and gastrointestinal motor function activators (specifically,acetylcholine agonists, dopamine receptor antagonists, dopamine D₂receptor antagonists, serotonin receptor agonists, opiate agonists, andChinese medicines (Liu Jun Zi Tang, Ban Xia Xie Xin Tang, and An ZhongSan), and gastrointestinal motor function suppressants (anticholinergicdrugs, muscarinic receptor antagonists, etc.), and the like. Suchdetermination can also be performed on a dyspeptic patient, and inparticular a patient with dyspepsia caused mainly by insufficientgastric motor functions (a patient with dysmotility-like dyspepsia), asa test subject. In this case, the pharmacotherapeutic effects onindividual dyspepsia patients can be determined, thereby making itpossible to select a suitable drug associated with gastrointestinalmotor functions (a gastrointestinal motor function improving agent,gastrointestinal motor function enhancer, or gastrointestinal motorfunction activator as mentioned above).

EXAMPLES

The present invention is described below with reference to Examples andTest Examples, which show production examples and evaluations of theproperties of preparations. However, the scope of the present inventionis not limited to these Examples and Test Examples.

Production Examples of Preparations Example 1

Twenty grams of ¹³C uracil and 380 g of D-mannitol (Mannit, a product ofKyowa Hakko Kogyo Co., Ltd.) were mixed, placed into a sample mill(KIIWG-1F, a product of Fuji Paudal Co., Ltd.), and mixed and pulverized(pulverization conditions: at a pulverization rotor speed of 12800 rpmand a sample feed motor speed of about 10 rpm, using a screen with 1-mmdiameter punched holes), to prepare a powder material. A 200-g quantityof the obtained powder material was weighed out into a speed kneader(NSK-150, a product of Okada Seiko Co., Ltd.), and 20 g of purifiedwater was added, followed by kneading. The resulting wet powder wasextruded through an extrusion granulator (Dome Gran DG-L, a product ofFuji Paudal Co., Ltd.) equipped with a dome-shaped die with 1-mmdiameter holes, and dried using an air-blow dryer (SPHH-200, a productof Espec Corp.) set at 60° C. Among the particles of the driedpreparation, those that passed through a sieve having a mesh size of1400 μm and did not pass through a sieve having a mesh size of 355 mwere obtained as a granular preparation containing 5 wt. % of ¹³Curacil.

The particle diameter of the thus obtained granular preparationcontaining 5 wt. % of ¹³C uracil was measured by a vibration sievemethod (specifically, using a Robot Shifter RPS-85 measurement apparatus(a product of Seishin Enterprise Co., Ltd.) at a vibration level of 5, ashift time of 5 minutes, and a pulse interval of 1 second). Table 1shows the results.

TABLE 1 Proportion Particle Diameter (wt. %) 1400 μm or more 2.09 Notless than 1000 μm and less than 1400 μm 7.29 Not less than 850 μm andless than 1000 μm 22.07 Not less than 710 μm and less than 850 μm 59.04Not less than 500 μm and less than 710 μm 8.99 Not less than 355 μm andless than 500 μm 0.09 Not less than 250 μm and less than 355 μm 0.00 Notless than 150 μm and less than 250 μm 0.09 Less than 150 μm 0.34 Total100.0

Comparative Example 1

Ten grams of ¹³C uracil and 190 g of D-mannitol (Mannit, a product ofKyowa Hakko Kogyo Co., Ltd.) were placed into a speed kneader (NSK-150,a product of Okada Seiko Co., Ltd.) and mixed, and then, withoutpulverization, 20 g of purified water was added, followed by kneading.Thereafter, granulation, drying, and particle size regulation by sievingwere carried out under the same conditions as in Example 1 to obtain agranular preparation containing 5 wt. % of ¹³C uracil. The particlediameter of the thus obtained granular preparation containing 5 wt. % of¹³C uracil was measured using the same method as in Example 1. Table 2shows the results.

TABLE 2 Proportion Particle Diameter (wt. %) 1400 μm or more 1.24 Notless than 1000 μm and less than 1400 μm 5.80 Not less than 850 μm andless than 1000 μm 30.39 Not less than 710 μm and less than 850 μm 54.87Not less than 500 μm and less than 710 μm 6.01 Not less than 355 μm andless than 500 μm 0.20 Not less than 250 μm and less than 355 μm 0.10 Notless than 150 μm and less than 250 μm 0.20 Less than 150 μm 1.19 Total100.0

Example 2

Twenty grams of ¹³C uracil and 180 g of D-mannitol (Mannit, a product ofKyowa Hakko Kogyo Co., Ltd.) were mixed, placed into a sample mill(KIIWG-1F, a product of Fuji Paudal Co., Ltd.), and mixed and pulverized(at a pulverization rotor speed of 12800 rpm and a sample feed motorspeed of about 10 rpm, using a screen with 1-mm diameter punched holes),to prepare a powder material. A 144-g quantity of the obtained powdermaterial was weighed out into a speed kneader (NSK-150, a product ofOkada Seiko Co., Ltd.), and 14.4 g of purified water was added, followedby kneading. The resulting wet powder was extruded through an extrusiongranulator (Dome Gran DG-L, a product of Fuji Paudal Co., Ltd.) equippedwith a dome-shaped die with 1-mm diameter holes, and dried using anair-blow dryer (SPHH-201, a product of Espec Corp.) set at 60° C. Amongthe particles of the dried preparation, those that passed through asieve having a mesh size of 1400 μm and did not pass through a sievehaving a mesh of 355 μm were obtained as a granular preparationcontaining 10 wt. % of ¹³C uracil.

Comparative Example 2

Twenty grams of ¹³C uracil and 180 g of D-mannitol (Mannit, a product ofKyowa Hakko Kogyo Co., Ltd.) were thoroughly mixed, and placed into aspeed kneader (NSK-150, a product of Okada Seiko Co., Ltd.). Twentygrams of purified water was added, followed by kneading. Subsequently,granulation, drying, and particle size regulation by sieving werecarried out under the same conditions as in Example 2 to obtain agranular preparation containing 10 wt. % of ¹³C uracil.

Comparative Example 3 Tablets

One hundred grams of ¹³C uracil, 60 g of lactose (a product of H.M.S),25 g of corn starch (a product of Nihon Shokuhin Kako Co., Ltd.), 10 gof crystalline cellulose (Ceolus PH301, a product of Asahi Kasei Co.),and 4 g of hydroxypropylcellulose (HPC-L fine powder, a product ofNippon Soda Co., Ltd.) were placed into a speed kneader (NSK-150, aproduct of Okada Seiko Co., Ltd.) and mixed. Forty grams of purifiedwater was then added, followed by kneading. Subsequently, the resultingkneaded powder was granulated using a speed mill (ND-02, a product ofOkada Seiko Co., Ltd.) equipped with a screen with 3-mm diameter punchedholes, and dried using an air-blow dryer (SPHH-200, a product of EspecCorp.) set at 70° C. The dried granules were sieved through a No. 16sieve for particle size regulation, and 1 g of magnesium stearate (aproduct of Taihei Chemical Industrial Co., Ltd.) was added to 199 g ofthe granules after particle size regulation to obtain granules fortablets. The granules for tablets were compressed into tablets eachweighing 200 mg using a single-punch tabletting machine (No. 2B, aproduct of Kikusui Seisakusho Ltd.) equipped with punches and dies witha diameter of 8 mm and rounded corners.

Example 3

Twenty grams of ¹³C uracil and 180 g of D-mannitol (Mannit, a product ofKyowa Hakko Kogyo Co., Ltd.) were thoroughly mixed, placed into a samplemill (SAM, a product of Nara Machinery Co., Ltd.), and mixed andpulverized (shape of grinding blades: pin-type; rotor speed: 4000 rpm,screen: a screen with 3-mm diameter punched holes), to obtain a powderpreparation.

Comparative Example 4

Twenty grams of ¹³C uracil was sieved through a No. 30 sieve to preparea powder preparation.

Comparative Example 5

Two hundred grams of ¹³C uracil was placed into a sample mill (SAM, aproduct of Nara Machinery Co., Ltd.) and pulverized under the sameconditions as in Example 3, to obtain a powder preparation.

Evaluation of Properties of Preparations Test Example 1 Particle SizeDistribution Measurement

The particle size distribution of the powder preparations of Example 3and Comparative Examples 4 and 5 was measured using a dry particle sizedistribution measuring apparatus (LDSA-1500A, a product of TohnichiComputer) under the following conditions: a focal distance of 100 mm, anumber of averaging processes of 10, an averaging interval of 5milliseconds, and an air pressure of 0.4 MPa. From the particle sizedistribution measured, the particle diameter at 10% (10% D), particlediameter at 50% (50% D), and particle diameter at 90% (90% D) werecalculated. Table 3 shows the results.

TABLE 3 10% D (μm) 50% D (μm) 90% D (μm) Ex. 3 5.74 14.95 56.58 Comp.Ex. 4 6.46 75.58 235.00 Comp. Ex. 5 6.01 52.60 260.57

As shown in Table 3, in the powder preparation of Comparative Example 4,which was obtained by sieving ¹³C uracil alone, and the powderpreparation of Comparative Example 5, which was obtained by pulverizing¹³C uracil alone, the particle diameter was not reduced, indicating aninsufficient pulverization effect, whereas in the powder preparation ofExample 3, which was obtained by mixing and pulverization, the particlediameter was reduced, demonstrating a sufficient pulverization effect.

Test Example 2 Evaluation of Solubility of Preparations

One hundred milliliters of tap water was added to each of 200-ml beakersat room temperature. Then, while stirring with a magnetic stirrer(RCN-7D, a product of EYELA) at 200 rpm, 2000 mg each of the granularpreparations of Example 1 and Comparative Example 1 was added to eachbeakers, and the time required for the preparations to dissolve wasmeasured by visual observation.

Further, three minutes after addition of the granular preparations, theamounts of undissolved residues of the preparations were visuallyevaluated.

Table 4 shows the results. As is apparent from the results, thepreparation of Comparative Example 1 took a long time to dissolve, and alarge amount of the preparation remained undissolved; whereas thepreparation of Example 1 dissolved in a short time, and only a smallamount of the preparation remained undissolved.

TABLE 4 Time Until Dissolution Undissoved Residue Ex. 1 1 min 10 secVery small amount Comp. Ex. 1 3 min or longer Larde amount

Test Example 3 Evaluation of Solubility of Preparations

Six tablets obtained in Comparative Example 3 were subjected to adisintegration test according to Japanese pharmacopoeia, 14th Edition,General Test Procedures, Disintegration Test. As a result, all thetablets had a disintegration time of 15 minutes or longer.

Test Example 4 Evaluation of Accuracy of Diagnosis of Pyridine MetabolicCapacity Disorder

After orally administering the preparations of Example 2 and ComparativeExample 2 to three healthy subjects (Subjects A, B, and C), air expiredfrom the subjects was collected over time and the ¹³C carbon dioxide gasconcentration in the expired air was measured using a GC-MS analyzer(ABCA-G, a product of Europa Scientific).

FIG. 2 shows the change in ¹³C carbon dioxide gas concentration in theexpired air after administration of the preparation of Example 2; andFIG. 3 shows the change in ¹³C carbon dioxide gas concentration afteradministration of the preparation of Comparative Example 2. In FIGS. 2and 3, the ordinate indicates A¹³C values (%), which are differencesbetween the δ ¹³C value (%) (¹³CO₂/¹²CO₂ concentration ratio) of theexpired air collected before administration of the preparation fordetermining pyrimidine metabolic capacity, and the δ ¹³C values (h) ofthe expired air collected at various periods of time afteradministration of the preparation. The abscissa indicates the periods(minutes) at which the expired air was collected after administration ofthe preparation. When the preparation of Comparative Example 2 wasadministered, the change in ¹³C carbon dioxide gas concentration wassmall in one of the three subjects, showing variation among the subjects(see FIG. 3). In contrast, when the preparation of Example 2 wasadministered to the same three subjects, the changes in ¹³C carbondioxide gas concentration in the subjects were similar to each other,showing only small variation among individuals. These resultsdemonstrate that a pyridine metabolic capacity disorder can be diagnosedrapidly, highly accurately, and with only small variation amongindividuals, by administering the preparation of Example 2 anddiagnosing a pyridine metabolic capacity disorder using, as an index,the ¹³C carbon dioxide gas concentration in the expired air collected 20to 30 minutes after administration of the preparation (see FIG. 2).

Test Example 5 Diagnosis of Gastric Emptying Capacity

The preparation of Example 1 was orally administered, at a dosecorresponding to 100 mg of 2-¹³C uracil, to human patients (20 cases)suspected of postoperative gastroparesis, within 20 days after thepatients had undergone stomach extraction operations. Air expired fromthe patients was collected 10, 20, 30, 40, 50, and 60 minutes afteradministration, and the ¹³CO₂ concentrations of the obtained expired airsamples, together with those of expired air samples (pre) collected inthe same manner before administration, were measured using GC/MS.Subsequently, the amount of change in ¹³CO₂ concentration (Δ ¹³C (%)) inthe expired air was calculated. FIG. 4 shows the results.

As shown in FIG. 4, the expired air test using the preparation of thepresent invention was able to classify the human patients (20 cases)into those with normal gastric emptying capacity (normal type: solidline), those with reduced gastric emptying capacity (delayed gastricemptying type: broken line), and those with insufficient gastricemptying capacity (insufficient type: dotted line). When the plasma2-¹³C uracil concentrations of these patients were measured 20 minutesafter administration of the preparation, a reduction in plasma 2-¹³Curacil concentration was observed in accordance with the gastricemptying capacity, in the patients with reduced gastric emptyingcapacity (delayed gastric emptying) and in the patients with gastricemptying capacity insufficiency. This demonstrates that the expired airtest using the oral preparation of the present invention effectivelyreflects the gastric emptying capacity.

1-18. (canceled)
 19. An oral preparation produced using a powdermaterial obtained by mixing component (a) and component (b) andpulverizing a resulting mixture, wherein a particle diameter at 50% ofthe powder material is 5 to 20 μm, (a) at least one isotope-labeledcompound selected from the group consisting of thymine, uracil and ametabolite thereof, in which at least one of carbon atoms, oxygen atoms,and nitrogen atoms is labeled with an isotope, and (b) at least onesugar alcohol selected from the group consisting of erythritol,mannitol, xylitol, sorbitol, maltitol, reducing paratinose and lactitol.20. An oral preparation according to claim 19, which contains thecomponent (a) in a proportion of 5 to 20 wt. %.
 21. An oral preparationaccording to claim 19, wherein the component (a) is isotope-labeleduracil.
 22. An oral preparation according to claim 19, wherein thecomponent (b) is mannitol.
 23. An oral preparation according to claim19, wherein the component (a) is isotope-labeled uracil and thecomponent (b) is mannitol.
 24. An oral preparation according to claim19, wherein the oral preparation is a solid preparation.
 25. An oralpreparation according to claim 24, wherein the solid preparation issubtle granules, granules, powders, tablets, capsules or pills.
 26. Anoral preparation according to claim 19, wherein the oral preparation isadministered at the same time as or immediately before or afteringestion of a test meal.
 27. A process for producing an oralpreparation, the process comprising the steps of: (1) producing a powdermaterial by mixing component (a) and component (B) and pulverizing aresulting mixture, wherein a particle diameter at 50% of the powdermaterial is 5 to 20 μm, (a) at least one isotope-labeled compoundselected from the group consisting of thymine, uracil and a metabolitethereof, in which at least one of carbon atoms, oxygen atoms, andnitrogen atoms is labeled with an isotope, and (b) at least one sugaralcohol selected from the group consisting of erythritol, mannitol,xylitol, sorbitol, maltitol, reducing paratinose and lactitol; and (2)formulating the powder material obtained in the above step (1) into apreparation.
 28. A process according to claim 27, wherein the oralpreparation contains the component (a) in a proportion of 5 to 20 wt. %.29. A process according to claim 27, wherein the component (a) isisotope-labeled uracil.
 30. A process according to claim 27, wherein thecomponent (b) is mannitol.
 31. A process according to claim 27, whereinthe component (a) is isotope-labeled uracil and the component (b) ismannitol.
 32. A process according to claim 27, wherein the oralpreparation is a solid preparation.
 33. A process according to claim 32,wherein the solid preparation is subtle granules, granules, powders,tablets, capsules or pills.
 34. A process according to claim 27, whereinthe oral preparation is administered at the same time as or immediatelybefore or after ingestion of a test meal.